It depends on the pH of the tap water and the contaminants therein.
Different areas may have very different stain reactions to tap water.  We
have very basic tap water and some things I did before will not work here
the same way.  The pH there was more acidic.  Pam Marcum

-----Original Message-----
From: amos brooks [mailto:atbrooks@snet.net]
Sent: Thursday, March 09, 2000 3:58 PM
To: DENISE BLAND-PIONTEK
Cc: histonet@pathology.swmed.edu
Subject: Re: Daily Digest


Hi,
    Three questions about this:
    1) When you are preparing the mucicarmine solution (diluting the stock)
are you using tap water or distilled? For some reason the tap water works
better, I dont know why. (If someone could enlighten me I'd be thrilled)
    2) Is there any autolysis in the epithelium of this stomach specimen?
Sometimes the goblet cells are the first to be affected.
    3) What part of the stomach are you staining? Some parts are less
reactive than others.
Amos Brooks

DENISE BLAND-PIONTEK wrote:

> RE:MUCICARMINE
>
> Dear Histonetters:
>      I have questions regarding a mucicarmine stain being done on distal
stomach.
> The stain has been done on different sections of stomach, distal to insure
mucin, but it is not turning out. We have checked the chemicals, expiration
dates (etc.). The same recipe and solutions have worked on other types of
tissue. Is there a specific reason why stomach is resistant to mucicarmine?
Any suggestions are appreciated.
>
> Thankyou,
> Denise Bland-Piontek, HTL(ASCP)
>
> >>> HistoNet Server <histonet@pathology.swmed.edu> 03/04/00 11:00PM >>>
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 03:45:17 -0600
> From: "Tony Henwood" <henwood@mail.one.net.au>
> Subject: Re:
>
> Dear Marianne,
> Would someone tell me, my gold chloride made in triple distilled water
> froze, is it still good to use?
>
> I can't imagine any staining technique that would be affected.
> Both water and gold chloride are unaffected by freezing.
> Why do you keep the solution in the fridge?
> Wouldn't room temp storage do?
>
> Regards, Tony
>
> Tony Henwood
> Senior Scientist
> Anatomical Pathology
> Royal Prince Alfred Hospital
> Sydney, AUSTRALIA
>
> http://www2.one.net.au/~henwood
> http://www.pathsearch.com/homepages/TonyHenwood/default.html
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 09:00:43 -0600
> From: "Tim Morken" <timcdc@hotmail.com>
> Subject: Re: cryostat decontamination
>
> Susan,
>
> You have discovered what I did last year. I did a literature search and
came
> up with nothing. Not one article on cryostat decontamination (at least
none
> that had documentation on why a certain method was used).
>
> Here are some methods I found that did not have any documentation to back
> them up as to effectiveness:
>
> In all cases you must defrost the cyrostat before decontaminating.
>
> 1) Wash out with 100% alcohol (be careful of flammable fumes)
>
> 2) wash with 10% formalin or Cidex (2.5 % glutaraldehyde) followed by
water,
> followed by 100% alcohol.
>
> This one is documented, but not published.
> 3) Put a dish of 37% formalin ("concentrated formalin", "formaldehyde") in
> the defrosted cryostat and leave overnight. This is from an unplublished
> study by a cryostat manufacturer and is the method recommended by Shandon
> for their cryostats.
>
> Textbooks will say to "decontaminate the cryostat" but either do not give
> any instructions on how to do so, or the instructions they give are not
> backed up with studies documenting effectiveness.
>
> No cryostat manufacturer, except Shandon, gives instructions on how to
> decontaminate a cryostat.
>
> I am working on a study to determine the need and effectiveness of
cryostat
> decontamination procedures, but that won't be done for awhile yet.
>
> It's not much, I know, but that's about it.
>
> Tim Morken, B.A., EMT(MSA), HTL(ASCP)
> Infectious Disease Pathology
> Centers for Disease Control
> MS-G32
> 1600 Clifton Rd.
> Atlanta, GA 30333
> USA
>
> email: tim9@cdc.gov
>        timcdc@hotmail.com
>
> Phone: (404) 639-3964
> FAX:  (404)639-3043
>
> - ----Original Message Follows----
> From: Susan McFarland <Susan.McFarland@lhsc.on.ca>
> To: HistoNet@Pathology.swmed.edu
> Subject: cryostat decontamination
> Date: Fri, 03 Mar 2000 14:01:16 -0500
>
> Hello there,
>
> I just did a search through the archives for procedures for disinfection
of
> cryostats and could find only questions, not answers. Does anyone have a
> protocol, with references for adequately disinfecting their cryostats they
> would be willing to share?  If it makes any difference with have both old
> Ames cryostats and newer Leica 1720s.
>
> I am in the process of trying to standardize protocols between two merged
> hospitals and find that neither protocol seems sufficient. I have nothing
to
> make a reference to in either case.
>
> Thanks very much,
>
> Sue McFarland,
> London Health Sciences Centre
> London, Canada
>
> ______________________________________________________
> Get Your Private, Free Email at http://www.hotmail.com
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 10:15:26 -0600
> From: "Molinari, Betsy" <BMolinari@heart.thi.tmc.edu>
> Subject: RE: LAB  FUMES
>
> Dear Sheila,
> Once a year we have our fume hoods checked and certified, for proper air
> exchange.  This is to meet GLP standards and for our own peace of mind.
> We are affiliated with a hospital  so the Maintanence Dept. takes care
> of it.
>
> > ----------
> > From:
> > sheila.cleveland@orst.edu[SMTP:sheila.cleveland@orst.edu]
> > Sent:         Friday, March 03, 2000 7:34 AM
> > To:   HistoNet Server
> > Subject:      LAB  FUMES
> >
> > Hi,
> > Is one sure they know if there lab is safe?  Sure hoods are
> > checked but what is safe??  I had a lab partner that became
> > chemicial sencitive and had to quiet her job.  If you can smell it, it
> >
> > must be bad, xylene, hemo de, etoh's, if they don't smell thats
> > even worse.  I try and work with a window open all year long, just to
> > give myself peace of mind.
> > sheila.cleveland@orst.edu
> >
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 11:00:21 -0600
> From: Jazz1522@aol.com
> Subject: Job openings
>
> There are a couple of openings for histotechs at US Labs in Irvine and Los
> Angeles, California.  If interested, fax resume to 949-450-0146 or call
949
> 450-0145 ext. 114,  for Human Resources.
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 13:00:11 -0600
> From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
> Subject: using 100% as decontaminant in cryostat
>
> I think the use of alcohol was discussed eons ago, as a decontaminating
agent
> in cryostat.  The question arose, 70% vs 95 or absolute alcohol, and 70%
was
> preferred for specific reasons, had something to do with suface tensions,
etc.
> hopefully someone will come forth with this again.
>
> Afer observing people inside of biohoods, cleaning down an area (wiping),
> they use 70% ethanol to reduce contamination by bacteria and some are
working
> with adenoviruses.  I figured if they get no contamination of their cell
> cultures, etc, with this type of cleaning, it would work in the cryostat
as
> well.
>
> To decontaminate, we defrost, wipe down with 70% several times on
absorbant
> towels, go to 100% to remove the residual water (cryostat is supposed to
be
> dry
> before putting it back together)  There are some wipe cloths available
now,
> from Current Technology that look promising, if people want to do a final
> wipedown after the 70% or even before that (when cryostat is warmed from
> defrosting).  There are other sources of wipe cloths, but I was always
afraid
> to put too much gunk in the cryostat, didn't want corrosion.
>
> We do wipe down areas in cryostat when it is running with 70% also, not a
lot,
> but enough to attract all the trimmings and after every use, if necessary.
>
> I think this in one of the tough areas to deal with, cryostat innards and
> protecting workers, and using formalin fumes just isn't acceptable.
>
> Out of curiosity, does everyone cryosection with gloves on these days in
the
> clinical setting?
>
> Gayle Callis
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 16:16:08 -0600
> From: amos brooks <atbrooks@snet.net>
> Subject: Re: cyclin D1
>
> Hi Vinnie,
>     We use Cyclin D1 frequently, on bone marrow biopsies. We use the DAKO
> antibody (M7155 ?) diluted to 1:50.
> The antigen is best retrieved using a high pH antigen retrieval. (also
from
> DAKO, I forget the pH) We use LSAB2
> for a detection kit. Don't even bother with Envision+, it doesn't work.
The
> incubation times following
> peroxidase blocking are 20 min primary, 10 min Link2, 10 min Label2 then
the
> chromogen you want. LSAB+ also
> works but we phased this one out after a bad lot problem.
>     The controls we use generally tend to be GI. We check each test we do
to
> determine if it can be used as a
> control. If it is positive with out ridiculous amounts of background then
we
> cut extra slides to hold us over
> until the next positive patient rolls along. If you are running low on
> controls, and a doctor asks for lymphoma
> markers on a GI case and not Cyclin D1 run one anyway just to check, that
is a
> good place to find them (Let the
> doctor see it of course, never file a slide a doctor hasn't seen!)
>     Just as a point of reference, I saw an incredible Cyclin D1 from a
> representative of SIGNET the other day.
> (made me drool all over the microscope what a mess :-) I think theirs blew
> ours out of the water. There was
> hardly any background staining, which is virtually impossible with this
quirky
> antibody. Unfortunately I dont
> know what their detection method was. :-(
> good luck
> Amos Brooks
>
> Vinnie Della Speranza wrote:
>
> > I would appreciate hearing from anyone who has this marker working
reliably
> to learn
> > 1. who's antibody you use
> > 2. your antigen recovery method
> > 3.  any caveats or tricks you might offer to provide better reliability
> > 4. what you are using for a control
> >
> > we have had some cases stain very well, others that the pathologist
insists
> should be positive that weren't.
> >
> > Vinnie Della Speranza
> > Manager for Anatomic Pathology Services
> > Medical University of South Carolina
> > 165 Ashley Avenue
> > Suite 309
> > Charleston, SC  29425
> > ph:  (843) 792-6353
> > fax: (843) 792-8974
> > email: Dellav@musc.edu
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 19:45:54 -0600
> From: JIGPEP@aol.com
> Subject: Re: tissue cassettes and breast processing
>
> Thanks to all who responded to my questions concerning the above topics.
It
> was greatly appreciated and I will be using some of the info. Carol,
> concerning the yellow baskets from the old Technicon that you have, I will
be
> glad to try them. I sent you an e-mail awhile ago, but perhaps it didn't
go
> through. We were having some trouble with aol then. You can respond thru
> histonet if you are still interested in releasing those items.
Thanks a
> million!!!       Jeanne Godine
>                                                           Pathology
> Consultants
>                                                           Lynchburg VA
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 20:45:30 -0600
> From: Victoria Baker <vbaker60@yahoo.com>
> Subject: Re: using 100% as decontaminant in cryostat
>
> Hi Gayle
>
> Where I worked previously they were rigid with this
> procedure.  Not using gloves could get a tech written
> up for not following procedure. Clinical settings
> leave the tech more unprotected from pathogens then
> those in research.  We have an idea of what we have to
> work with and the hazards it may impose.  Between TB
> resurfacing,AIDS and other "goodies", universal
> precaution is the rule.
>
> Vikki Baker
> American Health Foundation
> Valhalla, New York
>
> - --- Gayle Callis <uvsgc@msu.oscs.montana.edu> wrote:
> > I think the use of alcohol was discussed eons ago,
> > as a decontaminating agent
> > in cryostat.  The question arose, 70% vs 95 or
> > absolute alcohol, and 70% was
> > preferred for specific reasons, had something to do
> > with suface tensions, etc.
> > hopefully someone will come forth with this again.
> >
> > Afer observing people inside of biohoods, cleaning
> > down an area (wiping),
> > they use 70% ethanol to reduce contamination by
> > bacteria and some are working
> > with adenoviruses.  I figured if they get no
> > contamination of their cell
> > cultures, etc, with this type of cleaning, it would
> > work in the cryostat as
> > well.
> >
> > To decontaminate, we defrost, wipe down with 70%
> > several times on absorbant
> > towels, go to 100% to remove the residual water
> > (cryostat is supposed to be dry
> > before putting it back together)  There are some
> > wipe cloths available now,
> > from Current Technology that look promising, if
> > people want to do a final
> > wipedown after the 70% or even before that (when
> > cryostat is warmed from
> > defrosting).  There are other sources of wipe
> > cloths, but I was always afraid
> > to put too much gunk in the cryostat, didn't want
> > corrosion.
> >
> > We do wipe down areas in cryostat when it is running
> > with 70% also, not a lot,
> > but enough to attract all the trimmings and after
> > every use, if necessary.
> >
> > I think this in one of the tough areas to deal with,
> > cryostat innards and
> > protecting workers, and using formalin fumes just
> > isn't acceptable.
> >
> > Out of curiosity, does everyone cryosection with
> > gloves on these days in the
> > clinical setting?
> >
> > Gayle Callis
> >
> >
> >
> >
> __________________________________________________
> Do You Yahoo!?
> Talk to your friends online with Yahoo! Messenger.
> http://im.yahoo.com
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 22:15:28 -0600
> From: wbtyler@webtv.net (WB Tyler)
> Subject: Re: Azure B counterstain for melanocytic immunohistochemistry
>
> Someone requested information on this.
> This is the reference and the abstract.
> J Cutan Pathol 1991 Dec;18(6):436-9
> Immunoperoxidase technique modified by counterstain with azure B as a
> diagnostic aid in evaluating heavily pigmented melanocytic neoplasms.
> Kamino H, Tam ST
> Department of Dermatology, New York University Medical Center.
> Heavily-pigmented melanocytic neoplasms are difficult to evaluate on
> routine hematoxylin and eosin stained slides because pigmented
> melanocytes are difficult to distinguish from the numerous melanophages
> that are usually seen in the background of these lesions.
> Immunoperoxidase staining for S100 protein or HMB-45 antibody using
> diaminobenzidine (DAB) as chromogen, which forms a brown product, does
> not adequately distinguish melanocytes from melanophages. We modified
> this technique by replacing hematoxylin as the counterstain with azure
> B, which stains melanin green-blue. Thus, positive melanocytes appear
> brown while melanin granules in their cytoplasm are green-blue. However,
> negative melanophages only stain green-blue. This technique is useful in
> evaluating heavily pigmented melanocytic lesions such as malignant
> melanomas, melanosis of regressing malignant melanoma, residual
> malignant melanoma in areas of granulation tissue with melanophages,
> blue nevi, pigmented spindle cell variant of Spitz's nevi and combined
> nevi.
> PMID: 1723081, UI: 92129698
>
> William B. Tyler MD
> Department of Pathology
> Geisinger Medical Center
> Danville, PA
>
> ----------------------------------------------------------------------
>
> Date: 4 Mar 2000 22:36:55 -0600
> From: wbtyler@webtv.net (WB Tyler)
> Subject: Re: Spleen microanatomy
>
> Another reference for detailed spleen  histology is:
>
> Histology for Pathologists
> by Stephen S., Md. Sternberg (Editor)
>
> Hardcover - 1248 pages 2nd edition (January 1998)
> Lippincott Williams & Wilkins Publishers; ISBN: 0397517181
>
> This is available from Amazon.  $199
>
> William B. Tyler MD
> Dept of Pathology
> Geisinger Medical Center
> Danville, PA
>
> Here are the messages received yesterday!